An efficient method for isolation of high-quality RNA from mycelium of toxigenic fungi Fusarium sp.
- Authors: Stakheev А.А.1, Ryazantsev D.Y.1, Gabrielyan N.G.1, Poluboyarinova A.V.1, Taliansky М.E.1, Zavriev S.K.1
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Affiliations:
- Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
- Issue: Vol 59, No 2 (2025)
- Pages: 324-332
- Section: МЕТОДЫ
- URL: https://rjeid.com/0026-8984/article/view/682886
- DOI: https://doi.org/10.31857/S0026898425020138
- EDN: https://elibrary.ru/GFUSAT
- ID: 682886
Cite item
Abstract
Here, we describe a rapid and relatively cost-efficient method for the isolation and purification of RNA from mycelium of two species of plant-pathogenic fungi of Fusarium genus with different morphological and biochemical properties, F. graminearum and F. coffeatum. The method involves the use of guanidine hydrochloride-based buffer and spin columns from a commercial plasmid DNA extraction kit, and can be applied to both mycelia grown on nutrient agar media and liquid cultures of fungi. The yield of RNA isolated using the proposed protocol was 4–14 µg/100 mg of mycelium dry weight with RIN values up to 8.4. When optimizing the method, we propose to carry out a pre-lyophilization procedure, as well as the use of an RNAse inhibitor when isolating from cultures at late growth stages.
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About the authors
А. А. Stakheev
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Author for correspondence.
Email: stakheev.aa@gmail.com
Russian Federation, Moscow
D. Yu. Ryazantsev
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: stakheev.aa@gmail.com
Russian Federation, Moscow
N. G. Gabrielyan
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: stakheev.aa@gmail.com
Russian Federation, Moscow
A. V. Poluboyarinova
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: stakheev.aa@gmail.com
Russian Federation, Moscow
М. E. Taliansky
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: stakheev.aa@gmail.com
Russian Federation, Moscow
S. K. Zavriev
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: stakheev.aa@gmail.com
Russian Federation, Moscow
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